title: NanoSMGT: transfection of exogenous DNA on sex-sorted bovine sperm using nanopolymer.
abstract: The objective was to introduce exogenous DNA into commercially sex-sorted bovine sperm using nanopolymer for transfection. In the first experiment, the optimal concentration and ratio of linear-to-circular plasmid was determined for NanoSMGT in unsorted sperm. A second experiment was conducted to transfect exogenous DNA into sex-sorted sperm. Exogenous DNA uptake occurred in a dose-dependent manner (P < 0.05). The optimal amount of DNA was 10 μg/10(6) cells. The ratios of linear-to-circular plasmid do not influence the uptake by unsorted sperm cells and none of the tested treatments affected sperm motility and viability. Commercially sex-sorted bovine sperm were able to uptake exogenous DNA using nanopolymer; however, both X- and Y-sorted sperm had decreased DNA uptake in comparison to unsorted sperm (P < 0.05). Neither sperm motility nor viability were affected by nanotransfection. In conclusion, nanopolymer efficiently introduced exogenous DNA into commercially sex-sorted bovine sperm; we inferred that these sperm could be used for production of embryos of the desired sex, a technique named NanoSMGT.
authors.affiliation: Laboratório de Embriologia Molecular e Transgênese, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
authors.forename: V F
authors.initials: VF
authors.lastname: Campos
authors.forename: E R
authors.initials: ER
authors.lastname: Komninou
authors.forename: G
authors.initials: G
authors.lastname: Urtiaga
authors.forename: P M
authors.initials: PM
authors.lastname: de Leon
authors.forename: F K
authors.initials: FK
authors.lastname: Seixas
authors.forename: O A
authors.initials: OA
authors.lastname: Dellagostin
authors.forename: J C
authors.initials: JC
authors.lastname: Deschamps
authors.forename: T
authors.initials: T
authors.lastname: Collares
mesh_terms: D000818:Animals; D002417:Cattle / Q000235:genetics* / Q000502:physiology; D004247:DNA / Q000235:genetics*; D005822:Genetic Vectors; D008297:Male; D049329:Nanostructures; D012743:Sex Preselection; D013094:Spermatozoa / Q000502:physiology*; D014162:Transfection / Q000379:methods / Q000662:veterinary*
chemical_list: D004247:DNA